1.Establishment and optimization of a high-performance size-exclusion chromatography method for quantifying the classical swine fever virus E2 protein.
Xiaojuan ZHANG ; Bo YANG ; Gaoyuan XU ; Mingxing REN ; Ji TANG ; Hongshuo LIU ; Zhankui LIU ; Yafei LI ; Xiangru WANG
Chinese Journal of Biotechnology 2025;41(7):2774-2788
This study aims to establish a high-performance size-exclusion chromatography (HPSEC) method for determining the content of the classical swine fever virus (CSFV) E2 protein and screen the optimal stabilizer to enhance the stability of this protein. The optimal detection conditions were determined by optimizing the composition of the mobile phase, and characteristic chromatographic peaks were identified by SDS-PAGE and Western blotting. The specificity, repeatability, precision, linearity, limit of detection (LOD), and limit of quantitation (LOQ) of the method were assessed. The method established was used to determine the content of CSFV E2 protein antigen and vaccine. Differential scanning fluorimetry (DSF) was employed to screen the buffer system, pH, and salt ion concentrations, and sugar, amino acid, and alcohol stabilizers were further screened. The results showed that using a 200 mmol/L phosphate buffer provided the best column efficiency. An antigen-specific chromatographic peak appeared at the retention time of 18 min, which was identified as the CSFV E2 protein by SDS-PAGE and Western blotting. The method exhibited high specificity for detecting the CSFV E2 protein, with no absorbance peak observed in the blank control. The relative standard deviation (RSD) of the peak area for six repeated injections of the CSFV E2 protein was 0.74%, indicating good repeatability of the method. The RSD for repeated detection of two different concentrations of CSFV E2 protein samples by different operators at different time points was less than 2%, suggesting good intermediate precision of the method. The peak area of the CSFV E2 protein was linearly related to its concentration, with the regression equation showing R2 of 1.000. The LOD and LOQ of the method were 14.88 μg/mL and 29.75 μg/mL, respectively. Application of the developed method in the detection of three batches of CSFV E2 protein antigen and three batches of vaccine demonstrated results consistent with those from the bicinchoninic acid (BCA) assay, which meant that the method could accurately determine the content of CSFV E2 protein antigen and vaccine. The DSF method identified 50 mmol/L Tris-HCl at pH 8.0 as the optimal buffer, and the addition of sugar and alcohol stabilizers further improved the stability of the CSFV E2 protein. The HPSEC method established in this study is simple, fast, and exhibits good accuracy and repeatability, enabling precise measurement of the CSFV E2 protein content. It is expected to play a crucial role in the quality control of the CSFV E2 vaccine. Furthermore, the strategy for improving the CSFV E2 protein stability, identified through DSF screening, has significant implications for enhancing the stability of the CSFV E2 vaccine.
Classical Swine Fever Virus/chemistry*
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Chromatography, Gel/methods*
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Animals
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Swine
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Viral Envelope Proteins/immunology*
2.Comparison of efficacy of transpulmnary pressure and driving pressure in guiding lung-protective ventilation strategies for patients with acute respiratory distress syndrome
Yaoyao TANG ; Yan LI ; Wenli MENG ; Zhiyong WANG ; Mingxing FANG
Chinese Journal of Anesthesiology 2025;45(4):458-462
Objective:To compare the efficacy of transpulmonary pressure (Ptp) and driving pressure (DP) in guiding lung-protective ventilation strategies in patients with acute respiratory distress syndrome (ARDS).Methods:In this randomized controlled trial, 22 ARDS patients of either sex, aged 18-80 yr, with a body mass index of 18-30 kg/m 2, were divided into 2 groups ( n=11 each) using a random number table method: Ptp group and DP group. DP group received a tidal volume of 4-6 ml/kg with a plateau pressure<30 cmH 2O (1 cm H 2O=0.098 kPa) and driving pressure<16 cmH 2O. In Ptp group, the positive end-expiratory pressure (PEEP) and tidal volume were adjusted to maintain end-inspiratory Ptp <25 cmH 2O and end-expiratory Ptp 0-5 cmH 2O based on the setting mentioned in DP group. Oxygenation index, PEEP, plateau pressure, central venous pressure, heart rate, mean arterial pressure, parameters of liver and kidney function, mechanical ventilation duration, and the mortality in the intensive care unit were recorded on admission to the operating room and at 24 and 48 h after treatment. Results:Compared to DP group, significant changes were found in the main effects of oxygenation indices and time effects in Ptp group, and significant changes were found in the main effect of PEEP, with mean arterial pressure increasing ( P<0.05), and no significant differences were observed in the other parameters regarding group effects, time effects, or interactions in PEEP group ( P>0.05). Conclusions:Ptp provides a superior efficacy than DP in guiding lung-protective ventilation strategies for the patients with ARDS.
3.Research progress of novel three-dimensional echocardiographic rendering tools in structural heart disease
Alimu TEZHI ; Yuman LI ; Lin HE ; He LI ; Wenqian WU ; Yun YANG ; Huan WANG ; Mingxing XIE ; Lingyun FANG
Chinese Journal of Ultrasonography 2025;34(2):173-179
Recently developed three-dimensional echocardiography rendering technology,which includes transillumination imaging and tissue transparency imaging,is based on powerful artificial intelligence algorithm and employs unique visualization methods. This novel technology enables intuitive and realistic visualization of the heart's three-dimensional structure and blood flow images,providing richer and more accurate imaging information for evaluating cardiac anatomy and function. They demonstrate unique value in the diagnosis and treatment of structural heart diseases. This article reviews the applications and prospects of three-dimensional echocardiographic rendering techniques in structural heart disease.
4.Applications and mechanisms of the therapeutic low-intensity pulsed ultrasound in the immune cell modulation
Yingxin LI ; Tianshu LIU ; Qiaofeng JIN ; Xueke LI ; Jiani QIU ; Jingrong JIANG ; Mingxing XIE ; Jing WANG
Chinese Journal of Ultrasonography 2025;34(3):270-276
Low-intensity pulsed ultrasound(LIPUS),with its remarkable advantages of higher safety and better penetrability,has gradually become a novel method of physical adjuvant therapy. Previous studies have verified that LIPUS can modulate the immune response of different immune cells such as macrophage,T lymphocyte,and neutrophil by reducing the level of pro-inflammatory cytokines. Therefore,it plays a crucial role on acceleration of fracture healing,expedition of wound repair,and repairation of myocardial injury. The review summarizes the regulatory effects and potential mechanisms of LIPUS on abnormal immune cell responses triggered by various diseases.
5.Echocardiographic evaluation of pediatric mitral valve replacement:a single-center study
Linyue ZHANG ; Yuji XIE ; Zhaoli REN ; He LI ; Wenqian WU ; Li ZHANG ; Yuman LI ; Mingxing XIE
Chinese Journal of Ultrasonography 2025;34(1):17-25
Objective:To explore the application value of echocardiography in the management of pediatric mitral valve replacement.Methods:Thirty-three children who underwent mitral valve replacement at Union Hospital Tongji Medical College Huazhong University of Science and Technology from January 2009 to June 2023 were retrospectively analyzed. Clinical data and preoperative and postoperative echocardiographic data were collected. The differences in ultrasound parameters among preoperative,1-week postoperative,1-month postoperative and 1-year postoperative were compared.Results:Of the 33 children,there were 4 cases(12.12%)of mitral stenosis,33 cases(100%)of mitral regurgitation and 4 cases(12.12%)of mitral stenosis combined with regurgitation. Mechanical valve replacement was performed in 30 cases(90.91%)and bioprosthetic valve replacement in 3 cases(9.09%). Compared with the preoperative period,the left atrial internal diameter and left ventricular end-diastolic internal diameter were significantly reduced at 1-week postoperatively,1-month postoperatively,and 1-year postoperatively,respectively(all P <0.05).The left ventricular mass was significantly reduced and the left ventricular end-diastolic sphericity index was significantly increased(both P < 0.05)at 1-month postoperatively compared with preoperatively. The left ventricular ejection fraction was significantly higher at 1-year postoperatively compared with 1-week postoperatively( P <0.05). Conclusions:After pediatric mitral valve replacement,the left ventricular dimensions decrease,and cardiac function progressively recovers. Echocardiography is an essential tool for accurate preoperative assessment and postoperative follow-up in pediatric mitral valve replacement.
6.Effect of post-transplant diabetes mellitus on left ventricular longitudinal function and its prognostic value in heart transplant recipients
Yiwei ZHANG ; Yanting ZHANG ; Yuman LI ; Shuangshuang ZHU ; Wei SUN ; Yuji XIE ; Ye ZHU ; Mingxing XIE ; Li ZHANG
Chinese Journal of Ultrasonography 2025;34(5):377-382
Objective:To evaluate the left ventricular(LV)function in heart transplant(HTx)patients with post-transplant diabetes(PTDM),and to examine the relevance of PTDM and LV function to the patient's prognosis.Methods:Two hundred and thirteen adult HTx patients who underwent echocardiography at Union Hospital,Tongji Medical College,Huazhong University of Science and Technology between January 2018 and January 2022 were prospectively included. The patients were divided into PTDM group( n=86)and Non-PTDM group( n=127). LV function parameters were acquired using conventional and two-dimensional speckle-tracking echocardiography(2D-STE),and were compared between the two groups. The primary endpoints included all-cause mortality or transplant-related readmission. Results:Compared with Non-PTDM group,the LV mass of PTDM group was higher,the LV ejection fraction,LV global longitudinal strain(GLS),peak systolic global longitudinal strain rate,and early diastolic global longitudinal strain rate(dGLSr)were lower(all P<0.05). After a median follow-up period of 37.6(29.3)months,27 patients experienced clinical events. A multivariate analysis revealed that PTDM( HR=2.198,95% CI=1.018-4.743, P=0.045)and low GLS( HR=6.456,95% CI=2.889-14.426, P<0.001)were independent predictors of adverse clinical events after adjustment for dGLSr,body mass index and age. After subdividing the two groups into 4 subgroups by the cutoff value of GLS(16.5%),the prognosis was worst for HTx patients with PTDM and low GLS. Conclusions:HTx patients with PTDM have worse LV systolic and diastolic function than those without PTDM. Management of HTx patients with PTDM may be improved using GLS guidance.
7.Yttrium-90 resin microsphere selective internal radiation therapy on giant hepatocellular carcinoma in right lobe of liver: a case report
Nuoya LI ; Mingxing XU ; Jianliang XU
Chinese Journal of Clinical Medicine 2025;32(1):35-40
Primary liver cancer is a common malignant tumor in China. The current main treatment methods include surgical resection, liver transplantation, interventional therapy, radiofrequency ablation, chemotherapy, targeted therapy, and immunotherapy. Yttrium-90 microsphere selective internal radiation therapy (90Y-SIRT) represents a critical treatment option for converting unresectable intermediate to advanced hepatocellular carcinoma. This report details a case of a patient with a giant hepatocellular carcinoma in the right lobe treated with 90Y resin microsphere at the Third Affiliated Hospital of Sun Yat-sen University. Post-treatment, the tumor showed significant necrosis and reduction in size, with a marked decrease in alpha fetoprotein (AFP) levels and notable improvement in clinical symptoms. The case highlights the substantial efficacy of 90Y-SIRT in controlling tumor progression and enhancing patient quality of life.
8.Effects of Zhimu (Anemarrhena asphodeloides)-Huangbai (Phellodendron amurense) Medicinal Pair on Femoral Microstructure and Osteogenic-Adipogenic Differentiation in Ovariectomized Osteoporosis Model Rats
Chuncai LI ; Mingxing YUAN ; Jiawei LI ; Jing DENG ; Chongyang SHEN ; Yuan LIU
Journal of Traditional Chinese Medicine 2025;66(16):1704-1710
ObjectiveTo investigate the potential mechanisms of Zhimu (Anemarrhena asphodeloides)-Huangbai (Phellodendron amurense) medicinal pair in alleviating postmenopausal osteoporosis (PMOP). MethodsSixty unpregnant female SD rats were randomly divided into five groups, blank group, model group, low-dose Zhimu-Huangbai group, high-dose Zhimu-Huangbai group, and estradiol group, with 12 rats in each group. Except for the blank group, all other groups had their ovaries removed to create PMOP rat models, while the blank group only had the fat tissue around the ovaries removed. One week after the ovarian removal, the low-dose and high-dose Zhimu-Huangbai groups received concentrated solution of Zhimu and Huangbai with 1.8, 7.2 g/(kg·d) via gavage, the estradiol group received estradiol solution 0.09 mg/(kg·d) via gavage, and the blank group and the model group received 10 ml/(kg·d) of normal saline via gavage, once daily for 12 weeks. Before sampling, the body mass of the rats was recorded, and uterine tissue was taken to calculate the uterine index. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum were detected by ELISA; micro CT was used to examine the parameters of femoral microstructure, including bone volume/tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), bone mineral density (BMD), trabecular separation (Tb.Sp), and cortical bone area (Ct.Ar). HE staining was used to observe pathological changes in the femur; RT-qPCR was used to detect the mRNA expression of osteogenic-adipogenic differentiation-related factors in femoral tissue, including Runt-related transcription factor 2 (Runx2), bone morphogenetic protein 2 (BMP-2), peroxisome proliferator-activated receptor γ (PPARγ), chemerin and chemokine-like receptor 1 (CMKLR1). ResultsCompared with the blank group, the model group showed a significant increase in body mass, a significant decrease in the uterine index, BV/TV, Tb.N, Tb.Th and BMD, a significant increase in Tb.Sp, and serum IL-1β, IL-6, and TNF-α levels, a significant reduction of mRNA expression of Runx2 and BMP-2 in bone tissue, and a significant increased mRNA expression of PPARγ, chemerin, and CMKLR1 (P<0.01). HE staining revealed that the femoral tissue showed a reduction and sparsity of trabeculae, a significant enlargement of the medullary cavity, and a large number of fat cells. Compared to the model group, the low-dose, high-dose Zhimu-Huangbai groups, and estradiol group showed significant improvements in all the above-mentioned indicators (P<0.05 or P<0.01). HE staining revealed a significant increase in trabeculae, more organized arrangement, and a marked reduction in fat cells. Compared to low-dose Zhimu-Huangbai group, the high-dose Zhimu-Huangbai group exhibited a significant increase in the uterine index and BMD, and a significant reduction in body mass and PPARγ and Chemerin mRNA expression (P<0.05 or P<0.01). Compared to high-dose Zhimu-Huangbai group, the estradiol group showed a decrease in uterine index, BV/TV, Tb.N, Tb.Th, BMD, and BMP-2 mRNA expression, while the levels of IL-1β, TNF-α, and IL-6, as well as Tb.Sp and the mRNA expressions of PPARγ, chemerin, and CMKLR1 increased (P<0.05 or P<0.01). ConclusionThe Zhimu-Huangbai medicinal pair can alleviate PMOP bone loss, and its mechanism of action is related to reducing the levels of inflammatory factors, correcting the disorder of osteogenic-adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and promoting the differentiation of BMSCs into osteoblasts.
9.Development of DUS Test Guidelines for New Pinellia ternata
Xinyao LI ; Mingxing WANG ; Bingbing LIAO ; Changjie CHEN ; Xiufu WAN ; Lanping GUO ; Yuhuan MIAO ; Dahui LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):225-233
Pinellia ternata, belonging to the Pinellia genus within the Araceae family, is a medicinal plant due to its tubers. There are severe issues with unclear germplasm and mixed varieties in its cultivation, necessitating urgent new variety protection efforts. The distinctness, uniformity, and stability (DUS) testing of the plant variety is the basis for protecting new plant varieties, and the DUS test guidelines are the technical basis for DUS testing. To develop the DUS test guidelines for P. ternata, agronomic traits of 229 germplasm of P. ternata were observed and measured during its two growth stages over the years, and each character was graded and described. A total of 38 traits were selected as the test traits of the DUS test guideline for P. ternata. There were three plant traits, 19 leaf traits, six flower traits, two fruit traits, two tuber traits, five bulbil traits, and one ploidy trait. These traits could be divided into 22 quality characters, 12 quantitative characters, and four pseudo-quantitative characters, as well as seven groups, including plants, leaves, flowers, fruit, tubers, bulbils, and ploidy. By searching for standard traits, 10 standard varieties were ultimately determined. Preparing these guidelines will have great significance for reviewing and protecting P. ternata varieties, safeguarding breeders' rights, and promoting the development of the P. ternata industry.

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